| Small increases in pH enhance retroviral vector transduction efficiency of NIH-3T3 cells. | |
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MedLine Citation:
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PMID: 12573028 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Increases in pH between 7.1 and 7.7 increase the efficiency of polybrene (Pb)- and protamine sulfate (PS)-aided retroviral transduction of NIH-3T3 cells in a serum-lot-dependent manner. The increase in Pb-aided transduction efficiency at pH 7.7, relative to the value at pH 7.33, ranged from 13% to 49% for three serum lots. For a constant Moloney murine leukemia virus (MMLV) vector dilution at pH 7.33, three different serum lots resulted in absolute transduction efficiencies ranging from 29% to 53% using Pb. At the same vector dilution, PS-aided transduction was less effective on an absolute basis than Pb-aided transduction, but the benefit of elevated pH was more pronounced with PS. There was a similar enhancement with PS at elevated pH for a murine stem cell virus (MSCV) vector as for the MMLV vector. The benefit at pH 7.7 for PS-aided transduction was partially due to greater PS stability at elevated pH. Heat inactivating the serum supplement or adding protease inhibitors helped to stabilize PS. This increased the absolute transduction efficiency but decreased the relative benefit of elevated pH to a level similar to that for Pb-aided transduction. Incubating Pb with the vector at pH 7.1 for 10 min, prior to readjusting to pH 7.7 and transducing the cells, was sufficient to abrogate the beneficial effects of transduction at pH 7.7. In contrast, prior exposure of PS with vector at pH 7.1 did not affect subsequent transduction at pH 7.7. These results indicate that pH is an important variable in retroviral transduction and that the relative benefits of Pb or PS on retroviral vector transduction will vary with the pH, polymer addition method, and serum lot. |
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Authors:
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Tor W Jensen; Yong Chen; William M Miller |
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Publication Detail:
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Type: Evaluation Studies; Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S. |
Journal Detail:
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Title: Biotechnology progress Volume: 19 ISSN: 8756-7938 ISO Abbreviation: Biotechnol. Prog. Publication Date: 2003 Jan-Feb |
Date Detail:
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Created Date: 2003-02-07 Completed Date: 2003-10-07 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 8506292 Medline TA: Biotechnol Prog Country: United States |
Other Details:
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Languages: eng Pagination: 216-23 Citation Subset: IM |
Affiliation:
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Department of Chemical Engineering, Northwestern University, Evanston, Illinois 60208-3120, USA. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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3T3 Cells
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chemistry,
drug effects,
physiology* Animals Cell Culture Techniques / methods Cell Division / drug effects Culture Media / chemistry, pharmacology Genetic Vectors Hexadimethrine / pharmacology Hydrogen-Ion Concentration Mice Protamines / pharmacology Quality Control Reproducibility of Results Retroviridae / genetics* Sensitivity and Specificity Transduction, Genetic / methods* |
| Chemical | |
Reg. No./Substance:
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0/Culture Media; 0/Protamines; 9011-04-5/Hexadimethrine |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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